Affinity chromatography device including a fibrillated polymer membrane and a manifold including the same

Abstract

The present disclosure is directed to affinity chromatography devices comprising fibrillated polymer membranes containing inorganic particles having a spherical shape and a particle size distribution with a D90 / D10 of 3 or less. Blends or combinations of spherical inorganic particles may be utilized. The spherical inorganic particles have a nominal particle size of about 5 microns to about 20 microns. Affinity ligands can be bound to the spherical inorganic particles and / or the fibrillated polymer membrane. The affinity chromatography devices also include a fibrillated polymer membrane containing inorganic particles having a nominal particle size distribution of about 100 (×10 -12 cm 2 ) ~ approx. 500 (×10 -12 cm 2 ) hydraulic permeability. Additionally, the affinity chromatography device has a cycling durability of at least 100 cycles without exceeding an operating pressure of 0.3 MPa. Also disclosed are manifolds including multiple affinity chromatography devices in a parallel configuration, and multiple manifolds in a parallel configuration.

Description

Technical Field

[0001] Field The present disclosure generally relates to affinity chromatography, and more specifically to an affinity chromatography device that includes a fibrillated polymer membrane containing a blend of spherical particles therein, has a D90 / D10 of 3 or less for the particle size distribution, and thereby enables separation of target molecules from an aqueous mixture. A manifold including a plurality of affinity chromatography devices and a manifold in a parallel configuration are also disclosed.

Background Art

[0002] Background Chromatography methods are generally used to separate and / or purify target molecules such as proteins, nucleic acids, and polysaccharides from a mixture. Affinity chromatography specifically involves passing a mixture over a matrix having a ligand (i.e., a specific binding partner) specific for the target molecule to be bound. When contacted with the ligand, the target molecule binds to the matrix and is thus retained from the mixture. Affinity chromatography offers certain advantages over other types of chromatography. For example, affinity chromatography provides a purification method capable of separating a target protein from a mixture of the target protein and other biomolecules in a single step with high yield.

[0003] Despite the advantages of current affinity chromatography devices, there is a need in the art for a chromatography device that can be used with a shorter residence time than conventional devices, provides the same binding capacity or better binding capacity as currently available ones, and is reusable.

Summary of the Invention

[0004] Summary In one aspect (hereinafter referred to as "Aspect 1"), the affinity chromatography device includes a fluid inlet, a fluid outlet fluidly connected to the fluid inlet, a fibrillated polymer membrane disposed between the fluid inlet and the fluid outlet, the fibrillated polymer membrane having inorganic particles with a spherical shape and a nominal particle size of about 5 microns to about 20 microns therein, and a housing surrounding the fluid inlet, the fluid outlet, and the fibrillated polymer membrane. The particle size distribution has a D90 / D10 of 3 or less, and at least one of the fibrillated polymer membrane and the inorganic particles is covalently bonded to an affinity ligand that reversibly binds to a target molecule.

[0005] According to another aspect (hereinafter referred to as "Aspect 2"), in addition to Aspect 1, the target molecule is a protein, an antibody, a viral vector, and combinations thereof.

[0006] According to another aspect (hereinafter referred to as "Aspect 3"), in addition to Aspect 1 or Aspect 2, it has a water permeability of about 100 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ).

[0007] According to another aspect (hereinafter referred to as "Aspect 4"), in addition to any one of Aspects 1 to 3, the inorganic particles having a spherical shape are selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof.

[0008] According to another aspect (hereinafter referred to as "Aspect 5"), in addition to any one of Aspects 1 to 4, the fibrillated polymer membrane includes a stretched polytetrafluoroethylene membrane, a stretched modified polytetrafluoroethylene membrane, a stretched tetrafluoroethylene copolymer membrane, or an expanded (expanded, expanded, stretched, or foamed) polyethylene membrane.

[0009] According to another aspect (hereinafter referred to as "Aspect 6"), in addition to any one of Aspects 1 to 5, the fibrillated polymer membrane is a stretched polytetrafluoroethylene membrane.

[0010] According to another aspect ( "Aspect 7"), in addition to any one of Aspects 1 to 6, the affinity ligand is selected from Protein A, Protein G, Protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof.

[0011] According to another aspect ( "Aspect 8"), in addition to any one of Aspects 1 to 7, the inorganic particles at least include first inorganic particles having a spherical shape and a first nominal particle size, and second inorganic particles having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other.

[0012] According to another aspect ( "Aspect 9"), in addition to any one of Aspects 1 to 8, the nominal particle size is selected from about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof.

[0013] According to another aspect ( "Aspect 10"), in addition to any one of Aspects 1 to 9, the inorganic particles having a spherical shape include a blend of 10-micron spherical particles and 20-micron spherical particles, and the blend is from 10:90 to 90:10.

[0014] According to another aspect ( "Aspect 11"), in addition to any one of Aspects 1 to 9, the inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 10-micron spherical particles, and the blend is from 10:90 to 90:10.

[0015] According to another aspect ( "Aspect 12"), in addition to any one of Aspects 1 to 9, the inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 20-micron spherical particles, and the blend is from 10:90 to 90:10.

[0016] According to another aspect ( "Aspect 13"), in addition to any one of Aspects 1 to 12, it has a dynamic binding capacity (DBC) of at least 35 mg / ml with a residence time of 20 seconds.

[0017] According to another aspect ( "Aspect 14"), in addition to any one of Aspects 1 to 13, it has a cycle durability of at least 100 cycles at an operating pressure of less than 0.3 MPa.

[0018] According to another aspect ( "Aspect 15"), in addition to any one of Aspects 1 to 14, the fibrillated polymer film has a wound configuration.

[0019] According to another aspect ( "Aspect 16"), in addition to any one of Aspects 1 to 15, the fibrillated polymer film has a laminated configuration.

[0020] According to another aspect ( "Aspect 17"), in addition to any one of Aspects 1 to 16, the fibrillated polymer film has a wound configuration, a laminated configuration, and combinations thereof.

[0021] According to another aspect ( "Aspect 18"), in addition to any one of Aspects 1 to 17, the inner intermediate material is disposed around on the outer surface of the core, and the fibrillated polymer film is disposed around on the inner intermediate material.

[0022] According to another aspect ( "Aspect 19"), in addition to Aspect 18, the outer intermediate material is disposed around on the fibrillated polymer film.

[0023] According to another aspect ( "Aspect 20"), in addition to Aspect 18 or Aspect 19, the inner intermediate material and the outer intermediate material are selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous non-woven fabric material, and a porous woven fabric material.

[0024] According to another aspect ( "Aspect 21"), in addition to any one of Aspects 18 to 20, at least one of the inner intermediate material and the outer intermediate material is a polypropylene non-woven fabric material.

[0025] According to another aspect ( "Aspect 22"), an article according to any one of Aspects 18 to 21 is used for separating the target molecule from a fluid flow.

[0026] In one aspect (the "Aspect 23"), the manifold includes at least two affinity chromatography devices according to any one of Aspects 1 to 21 arranged in a parallel configuration.

[0027] According to another aspect (the "Aspect 24"), in addition to Aspect 23, the manifold is enclosed within a housing.

[0028] In one aspect (the "Aspect 25"), the article includes a first manifold and a second manifold in a parallel configuration, and each of the first manifold and the second manifold includes a plurality of affinity chromatography devices according to any one of Aspects 1 to 21.

[0029] In another aspect (the "Aspect 26"), in addition to Aspect 25, the first manifold and the second manifold are enclosed within a housing.

[0030] In one aspect (the "Aspect 27"), the article includes a core disposed at the center, a fibrillated polymer membrane containing spherical inorganic particles having a spherical shape and a nominal particle size of about 5 microns to about 20 microns wound around the core, a housing member surrounding both the core and the fibrillated polymer membrane, a first end cap disposed at a first end of the housing member, and a second end cap disposed at a second end of the housing member, the particle size distribution having a D90 / D10 of less than 3, and at least one of the fibrillated polymer membrane and the spherical inorganic particles covalently bonded to an affinity ligand that reversibly binds to a target molecule.

[0031] According to another aspect (the "Aspect 28"), in addition to Aspect 27, the target molecule is a protein, an antibody, a viral vector, and combinations thereof.

[0032] According to another aspect (the "Aspect 29"), in addition to Aspect 27 or Aspect 28, about 100 (×10-12 cm 2 ) to about 500 (×10 -12 cm 2 ) and has a water permeability.

[0033] According to another aspect ( "Aspect 30"), in addition to any one of Aspects 27 to 29, the inorganic particles having a spherical shape are selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof.

[0034] According to another aspect ( "Aspect 31"), in addition to any one of Aspects 27 to 30, the fibrillated polymer film includes at least one of a stretched polytetrafluoroethylene film, a stretched modified polytetrafluoroethylene film, a stretched tetrafluoroethylene copolymer film, or an expanded polyethylene film.

[0035] According to another aspect ( "Aspect 32"), in addition to any one of Aspects 27 to 31, the fibrillated polymer film is a stretched polytetrafluoroethylene film.

[0036] According to another aspect ( "Aspect 33"), in addition to any one of Aspects 27 to 32, the affinity ligand is selected from Protein A, Protein G, Protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof.

[0037] According to another aspect ( "Aspect 34"), in addition to any one of Aspects 27 to 33, the inorganic particles include at least a first inorganic particle having a spherical shape and a first nominal particle size, and a second inorganic particle having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other.

[0038] According to another aspect ( "Aspect 35"), in addition to any one of Aspects 37 to 34, the nominal particle size is selected from about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof.

[0039] According to another aspect ( "Aspect 36"), in addition to any one of Aspects 27 to 34, the inorganic particles having a spherical shape include a blend of 10-micron spherical particles and 20-micron spherical particles, and the blend is 10:90 to 90:10.

[0040] According to another aspect ( "Aspect 37"), in addition to any one of Aspects 27 to 34, the inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 10-micron spherical particles, and the blend is 10:90 to 90:10.

[0041] According to another aspect ( "Aspect 38"), in addition to any one of Aspects 27 to 34, the inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 20-micron spherical particles, and the blend is 10:90 to 90:10.

[0042] According to another aspect ( "Aspect 39"), in addition to any one of Aspects 27 to 38, it has a dynamic binding capacity (DBC) of at least 35 mg / ml with a residence time of 20 seconds.

[0043] According to another aspect ( "Aspect 40"), in addition to any one of Aspects 27 to 39, it has a cycle durability of at least 100 cycles and an operating pressure of less than 0.3 MPa.

[0044] According to another aspect ( "Aspect 41"), in addition to any one of Aspects 27 to 40, the inner intermediate material is disposed around on the outer surface of the core, and the fibrillated polymer membrane is disposed around on the inner intermediate material.

[0045] According to another aspect ( "Aspect 42"), in addition to any one of Aspects 27 to 41, it includes an outer intermediate material disposed around on the fibrillated polymer membrane.

[0046] According to another aspect ( "Aspect 43"), in addition to Aspect 42, the inner intermediate material and the outer intermediate material are selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous non-woven material, and a porous woven material.

[0047] According to another aspect ( "Aspect 44"), in addition to Aspect 43, at least one of the inner intermediate material and the outer intermediate material is a polypropylene non-woven material.

[0048] According to one aspect ( "Aspect 45"), the manifold includes at least two affinity chromatography devices according to any one of Aspects 27 to 44 arranged in a parallel configuration.

[0049] According to another aspect ( "Aspect 46"), in addition to Aspect 45, the manifold is enclosed within a housing.

[0050] According to one aspect ( "Aspect 47"), the device includes a first manifold and a second manifold in a parallel configuration, and each of the first manifold and the second manifold includes at least two affinity chromatography devices according to any one of Aspects 27 to 44.

[0051] According to another aspect ( "Aspect 48"), in addition to Aspect 47, the first manifold and the second manifold are enclosed within a housing.

[0052] In one aspect (hereinafter referred to as "Aspect 49"), the affinity chromatography device includes a housing, an inlet that enables fluid flow into the housing, a first flow distributor and a second flow distributor, where the first flow distributor and the second flow distributor are disposed at both ends of the housing, an outlet that enables fluid flow out of the housing, and a laminated membrane assembly disposed within the housing between the fluid inlet and the fluid outlet. Here, the laminated membrane assembly includes two or more fibrillated polymer membranes in a laminated configuration, and the fibrillated polymer membranes contain internally a blend of spherical inorganic particles having a nominal particle size of about 5 microns to about 20 microns. The particle size distribution has a D90 / D10 of less than 3, and at least one of the fibrillated polymer membrane and the spherical inorganic particles is covalently bonded to an affinity ligand that reversibly binds to a target molecule.

[0053] According to another aspect (hereinafter referred to as "Aspect 50"), in addition to Aspect 49, it includes a first flow distributor and a second flow distributor, where the first flow distributor and the second flow distributor are disposed at both ends of the housing member.

[0054] According to another aspect (hereinafter referred to as "Aspect 51"), in addition to Aspect 49 or Aspect 50, the target molecule is a protein, an antibody, a viral vector, and combinations thereof.

[0055] According to another aspect (hereinafter referred to as "Aspect 52"), in addition to any one of Aspects 49 to 51, it has a water permeability of about 100 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ).

[0056] According to another aspect (hereinafter referred to as "Aspect 53"), in addition to any one of Aspects 49 to 52, the spherical inorganic particles are selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof.

[0057] According to another aspect ( "Aspect 54"), in addition to any one of Aspects 49 to 53, the fibrillated polymer film includes a stretched polytetrafluoroethylene film, a stretched modified polytetrafluoroethylene film, a stretched tetrafluoroethylene copolymer film, or an expanded polyethylene film.

[0058] According to another aspect ( "Aspect 55"), in addition to any one of Aspects 49 to 54, the fibrillated polymer film is a stretched polytetrafluoroethylene film.

[0059] According to another aspect ( "Aspect 56"), in addition to any one of Aspects 49 to 55, the affinity ligand is selected from Protein A, Protein G, Protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof.

[0060] According to another aspect ( "Aspect 57"), in addition to any one of Aspects 49 to 56, the inorganic particles include at least a first inorganic particle having a spherical shape and a first nominal particle size, and a second inorganic particle having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other.

[0061] According to another aspect ( "Aspect 58"), in addition to any one of Aspects 49 to 57, the nominal particle size is selected from about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof.

[0062] According to another aspect ( "Aspect 59"), in addition to any one of Aspects 49 to 58, the inorganic particles having a spherical shape include a blend of 10 - micron spherical particles and 20 - micron spherical particles, and the blend is from 90:10 to 10:90.

[0063] According to another aspect ( "Aspect 60"), in addition to any one of Aspects 49 to 58, the inorganic particles having a spherical shape include a blend of 5 - micron spherical particles and 10 - micron spherical particles, and the blend is from 10:90 to 90:10.

[0064] According to another aspect (hereinafter referred to as "Aspect 61"), in addition to any one of Aspects 49 to 58, the inorganic particles having a spherical shape include a blend of spherical particles of 5 microns and spherical particles of 20 microns, and the blend is in the range of 10:90 to 90:10.

[0065] According to another aspect (hereinafter referred to as "Aspect 62"), in addition to any one of Aspects 49 to 61, it has a dynamic binding capacity (DBC) of at least 35 mg / ml with a residence time of 20 seconds.

[0066] According to another aspect (hereinafter referred to as "Aspect 63"), in addition to any one of Aspects 49 to 62, it has a cycle durability of at least 100 cycles and an operating pressure of less than 0.3 MPa.

[0067] According to another aspect (hereinafter referred to as "Aspect 64"), in addition to any one of Aspects 49 to 63, at least one first intermediate material is disposed on the first side of the laminated film assembly, and a second intermediate material is disposed on the second side of the laminated film assembly, and the second side is opposite to the first side.

[0068] According to another aspect (hereinafter referred to as "Aspect 65"), in addition to Aspect 64, the first intermediate material and the second intermediate material are selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous nonwoven material, and a porous woven material.

[0069] According to another aspect (hereinafter referred to as "Aspect 66"), in addition to Aspect 65, at least one of the first intermediate material and the second intermediate material is a polypropylene nonwoven material.

[0070] In one aspect (hereinafter referred to as "Aspect 67"), the manifold includes at least two affinity chromatography devices according to any one of Aspects 49 to 66 arranged in a parallel configuration.

[0071] According to another aspect (hereinafter referred to as "Aspect 68"), in addition to Aspect 67, the manifold is enclosed within a housing.

[0072] In one aspect ( "Aspect 69"), the device includes a first manifold and a second manifold in a parallel configuration, and each of the first manifold and the second manifold includes an affinity chromatography device according to any one of at least two aspects 49-66.

[0073] According to another aspect ( "Aspect 70"), in addition to Aspect 69, the first manifold and the second manifold are enclosed within a housing.

[0074] In another aspect ( "Aspect 71"), the diagnostic article includes a fibrillated polymer membrane containing spherical and inorganic particles having a nominal particle size of about 5 microns to about 20 microns therein, the particle size distribution has a D90 / D10 of 3 or less, and at least one of the fibrillated polymer membrane and the inorganic particles is covalently bonded to a ligand that reversibly binds to a target substance in a biological fluid.

[0075] According to another aspect ( "Aspect 72"), in addition to Aspect 71, it includes a fluid inlet and a fluid outlet fluidly connected to the fluid inlet.

[0076] According to another aspect ( "Aspect 73"), in addition to Aspect 71 or Aspect 72, it includes a housing member surrounding the fluid inlet, the fluid outlet, and the fibrillated polymer membrane. BRIEF DESCRIPTION OF THE DRAWINGS

[0077] Brief Description of the Drawings The accompanying drawings are included to provide a further understanding of the present disclosure, are incorporated herein, form a part of this specification, illustrate embodiments, and together with the description serve to explain the principles of the present disclosure.

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DETAILED DESCRIPTION OF THE INVENTION

[0093] Detailed Description Those skilled in the art will readily understand that the various aspects of the present disclosure can be realized by any number of methods and apparatuses configured to perform the intended functions. It should also be noted that the accompanying figures referred to in this specification are not necessarily drawn to scale and may be exaggerated to show the various aspects of the present disclosure, and in that regard, the figures should not be construed as limiting. As used herein, the term "on" is to be understood to mean that an element such as a polymer film is directly above another element or that intervening elements may also be present.

[0094] It should be understood that the terms "spherical particle", "spherical inorganic particle" and "inorganic particle having a spherical shape" are interchangeable in this specification. Further, the term "helical wound membrane assembly" is intended to include both a fibrillated polymer membrane alone and a combination of a fibrillated polymer membrane and an intermediate nonwoven material. Further, the term "laminated membrane assembly" is intended to include both a fibrillated polymer membrane alone and a combination of a fibrillated polymer membrane and one or more intermediate materials. Further, the terms "affinity chromatography device" and "chromatography device" may be used interchangeably in this specification.

[0095] The present disclosure is directed to an affinity chromatography device for separating a target molecule from an aqueous mixture containing the target molecule. Target molecules include, but are not limited to, proteins, antibodies, viral vectors, and combinations thereof. In some embodiments, the present disclosure is directed to a diagnostic device for separating a target disease from a biological sample. The chromatography device and the diagnostic device include a fibrillated polymer membrane containing a blend of inorganic particles having a particle size distribution that is spherical and has a D90 / D10 of 3 or less. In some embodiments, a blend or combination of spherical inorganic particles of various sizes is utilized. The nominal particle size of the spherical inorganic particles is from about 5 microns to about 20 microns. An affinity ligand can be bound to the spherical inorganic particles and / or the fibrillated polymer membrane. Further, the chromatography device has a dynamic binding capacity (DBC) of greater than 40 mg / ml at 10% breakthrough with a residence time of 20 seconds. Further, the affinity chromatography device has a cycle durability of at least 100 cycles without the operating pressure exceeding 0.3 MPa. As used herein, the term "about" is understood to represent + / - 10% of the specified unit of measurement.

[0096] Referring to FIGS. 1 and 2, a wound chromatography device 100 is shown. In forming the chromatography device 100, at least one fibrillated polymer membrane containing spherical inorganic particles therein is wound around a cylindrical core 150. Fibrillation, as used herein, refers to including fibrils within the polymer membrane, for example, a microstructure characterized by nodes interconnected by fibrils and having a microstructure where voids are the spaces between the nodes and fibrils. In some embodiments, at least one inner intermediate material 200 can be circumferentially disposed (e.g., wound) around the core 150 to a desired width or a pre-specified amount. Next, the fibril polymer membrane 210 containing spherical inorganic particles therein is wound around the core 150 on the inner intermediate material 200 to a desired width or a pre-specified amount, and an outer layer of at least one outer intermediate material 220 is circumferentially disposed (e.g., wound) around the fibrillated polymer membrane 210 to a desired width or a pre-specified amount. As used herein, the combination of the inner intermediate material 200, the fibrillated polymer membrane 210, and the outer intermediate material 220 is referred to as a "wound membrane assembly". The "wound membrane assembly" can include, in some embodiments, a fibrillated polymer membrane and an inner intermediate material wound around a core, a fibrillated polymer membrane and an outer intermediate material, and any combination of polymers and / or polymers and intermediate materials. The cylindrical core 150 can have a hollow or solid interior. In either case, the core 150 includes a solid outer wall such that an aqueous mixture flowing through the chromatography device 100 flows through an inner flow path formed from the inner intermediate material, which will be discussed in detail below. The use of a hollow core 150 reduces the amount of material used to form the core 150, reduces the weight of the device 100, and reduces the manufacturing cost.

[0097] The membrane assembly 110 and the central core 150 can be disposed within the housing 50. In some embodiments, the housing 50 is cylindrical. In the embodiments shown in FIGS. 1 and 2, the outer intermediate material 220 forms the outer flow path 130 and the inner intermediate material 200 forms the inner flow path 140. It should be understood that the intermediate materials 200, 220 may be different or the same in the embodiments described herein. Further, two or more intermediate materials can be used to form one or both of the outer flow path 130 and the inner flow path 140. In use, the aqueous mixture flows into an inlet 80 disposed within an inlet cap 60, the mixture flows over a dispenser cap 65 and is directed into the outer flow path 130 formed by the outer intermediate material 220. The dispenser cap 65 directs the aqueous mixture 90 degrees from the supply direction towards the outer flow path 130 (i.e., the intermediate material 220). This change in direction facilitates a more uniform flow of the aqueous mixture into the outer flow path 130. The outer intermediate layer 220 forming the outer flow path 130 is positioned between the housing 50 and the wound polymer membrane 210. The dispenser cap 65 may be a polyolefin or may be coated with a polyolefin. It should be understood that the aqueous mixture flows along the outer flow path gap 165 and connects to the inner flow path 130.

[0098] The aqueous mixture flows through the outer flow path 130 (i.e., the outer intermediate material 220) and across the wound polymer membrane 210 in a normal direction (e.g., normal flow). When the aqueous mixture passes through the outer flow path 130 (i.e., the outer intermediate material 220) and across the wound polymer membrane 210 in a normal flow, the affinity ligand reversibly binds to the target protein, thereby effectively removing it from the aqueous mixture. The aqueous mixture then enters the inner flow path 140 (i.e., the inner intermediate material 200) located between the solid outer wall of the central core 150 and the wound polymer membrane 210.

[0099] Next, the aqueous mixture is redirected at the bottom of the internal flow path 140 by the outlet cap 75. The aqueous mixture then exits the chromatography device 100 through an outlet 85 located within the outlet cap 75. It should be understood that the diameter and / or height of the central core 150 (and / or the width and / or height of the fibrillated polymer membrane and / or the intermediate material) can be adjusted to achieve a larger volume without adversely affecting the performance of the device. Further, as is known to those skilled in the art, for example, a target protein can be removed from an affinity ligand by passing a fluid having a lower pH through the chromatography device.

[0100] The intermediate materials 200, 220, and 40 are not particularly limited as long as the aqueous mixture can flow therethrough. Some non-limiting examples of suitable intermediate materials include, but are not limited to, a porous fluoropolymer film or a porous non-fluoropolymer film (e.g., a porous polypropylene or other porous polyolefin film), a porous nonwoven material, or a porous woven material. In some embodiments, the wound membrane assembly includes an integrated inlet end cap 60 at one end of the core 150 and an integrated outlet end cap 75 at the opposite end of the core 150, forming an integrated reusable chromatography device.

[0101] The total number of fibrillated polymer membrane layers present in the wound membrane assembly is not particularly limited and depends on the desired end use and / or the desired mass transport flux within the membrane assembly. The wound membrane assembly can include a total of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 (or more) total polymer membrane layers. It should be understood that hundreds or even thousands of polymer membrane layers can be present in the laminated membrane assembly. Further, the fibrillated polymer membranes present in the wound membrane assembly can have a single layer thickness of from about 1 micron to about 10,000 microns, from about 100 microns to about 5,000 microns, up to about 500 microns to about 3,000 microns, or from about 650 microns to about 1,000 microns. As used herein, the term "thickness" is in the direction of the fibrillated polymer membrane perpendicular to the length region of the fibrillated polymer membrane.

[0102] The functionality of the chromatography device 300 shown in FIG. 3 is substantially similar to that of the chromatography device 100 shown in FIG. 2. For example, an aqueous mixture is introduced into the chromatography device 300 in the direction of arrow 62 through an inlet 80 disposed within an inlet cap 60. The aqueous mixture is directed from the supply direction by a distribution cap 65. For ease of explanation, the aqueous mixture can flow in the direction indicated by arrow 55 towards an outer flow path 130 (which can be formed from an outer intermediate material 220). The aqueous mixture flows along an outer flow path gap 165 and connects to an inner flow path 140, where it flows in the direction of arrow 30. The aqueous mixture flows in a normal direction (e.g., a normal flow) across the wound polymer membrane 210 from the inner flow path 130 to the inner flow path 140 as indicated by arrow 70. When the aqueous mixture passes through the wound polymer membrane 210, the affinity ligand reversibly binds to the target molecule. It should be understood that the inner flow path 140 can be formed from an inner intermediate material 200.

[0103] The aqueous mixture that does not contain the target molecule flows down the inner flow path 140 in the direction indicated by arrow 40. The aqueous mixture is redirected at the bottom of the inner flow path 140 towards the central portion of the chromatography device 300 as shown by arrow 52. The aqueous mixture that does not contain the target molecule flows out of the chromatography device 300 in the direction of arrow 45 through an outlet 85 disposed within the outlet end cap 75.

[0104] In other embodiments, as generally shown in FIG. 4, the chromatography device 200 includes a fibrillated polymer membrane configured as individual disks 240 laminated on top of each other to form a laminated membrane assembly 220. The fibrillated polymer membranes 240 can be arranged in a laminated configuration simply by laminating the fibrillated polymer membrane disks 240 on top of each other. Alternatively, the fibrillated polymer membrane disks 240 can be laminated and subsequently laminated together by heat and / or pressure or other conventional methods. It should be understood that the laminated membrane assembly 240 described herein is with respect to fibrillated polymer membrane disks for ease of explanation. Fibrillated polymer membranes formed in geometric and / or non-geometric shapes are considered to be within the scope of the present disclosure.

[0105] The chromatography device 200 includes at least one upper intermediate material 260 disposed on top of the laminated membrane assembly and at least one lower intermediate material 280 disposed below the laminated membrane assembly 220. The upper intermediate material 260 and the lower intermediate material 280 may be the same or different from each other. Similar to the wound membrane assembly described above, the intermediate materials 260, 280 used to form the laminated membrane assembly 220 are not particularly limited as long as the aqueous mixture can flow through them. Non-limiting examples of suitable intermediate materials include, but are not limited to, porous fluoropolymer films or porous non-fluoropolymer films (e.g., porous polypropylene or other porous polyolefin films), porous non-woven materials or porous woven materials.

[0106] The laminated film assembly 220 can be disposed within a housing 250 having an inlet cap 265 and an outlet end cap 275 disposed at opposite ends of the housing 250. In some embodiments, the housing 250 is cylindrical, but any geometry capable of housing the laminated film assembly and achieving the desired dynamic binding capacity is considered within the scope of the present disclosure. In some embodiments, the intermediate materials 260, 280, the housing 250, the inlet cap 265, and the outlet cap 275 can be formed from a thermoplastic polymer such as polypropylene, polyethylene, or other polyolefin. Alternatively, one or both of the intermediate materials 260, 280 can be formed from an inorganic or metallic material, provided that the porous intermediate materials 260, 280 do not interfere with the operation of the chromatography device.

[0107] The fibrillated polymer membrane 240 within the laminated film assembly 220 can be adhered to the housing 250 at the inner wall of the housing 250 via any conventional process (e.g., melt sealing or use of a sealant) that prevents flow between the periphery of the fibrillated polymer membrane 240 and the housing 250. The inlet cap 265 and the outlet cap 275 can be sealed to the housing 250 by a similar or identical process. The inlet cap and the outlet caps 265, 275 each include an inlet 280 and an outlet 285, respectively, and allow for the flow of an aqueous mixture through the affinity chromatography device 200. Specifically, the inlet cap 265 allows for the fluid flow of the aqueous mixture into the housing 250, and the outlet cap 285 allows for the fluid flow of the aqueous mixture out of the housing 250. During use, the aqueous mixture sequentially flows through the intermediate material 260, through the fibrillated polymer membrane 240 forming the laminated film assembly 220, and through the intermediate material 280. As the aqueous mixture passes through the chromatography device 200, the affinity ligand reversibly binds to the target molecule, thereby effectively removing it from the aqueous mixture. The target molecule can be removed from the affinity ligand, for example, by passing a fluid having a lower pH through the device, as is known to those skilled in the art.

[0108] The total number of fibrillated polymer membranes present in the laminate film assembly is not particularly limited and depends on the desired end use and / or the desired mass transport flux within the membrane assembly. The laminate film assembly can include a total of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 (or more) polymer membranes. It should be understood that there can be hundreds or even thousands of polymer membranes in the laminate film assembly. Further, the fibrillated polymer membranes present in the laminate film assembly can have a thickness of from about 1 micron to about 10,000 microns, from about 100 microns to about 5,000 microns, from about 500 microns to about 3,000 microns, or from about 650 microns to about 1,000 microns. As used herein, the term "thickness" is in the direction of the fibrillated polymer membrane perpendicular to the length region of the fibrillated polymer membrane.

[0109] The fibrillated polymer membranes in both the wound film assembly and the laminate film assembly include spherical inorganic particles or particles having a spherical configuration. As used herein, the term "spherical" is intended to mean that the inorganic particles have a circular or substantially circular shape in which the distance from the center of the inorganic particle to the outer edge of the particle at any point is the same distance or substantially the same distance. In some embodiments, the spherical inorganic particles have a particle size distribution having a D90 / D10 of 3 or less, 2.5 or less, 2 or less, 1.5 or less, or 1 or less. The spherical inorganic particles can have a nominal particle size of about 5 microns, about 10 microns, about 15 microns, about 20 microns, and can be combinations and blends thereof. In some embodiments, the spherical inorganic particles are polydisperse.

[0110] In some embodiments, the fibrillated polymer membrane contains more than one nominal particle size and / or more than one type of spherical inorganic particles within the fibrillated polymer membrane. The fibrillated polymer membrane can contain about 10 wt% fibrillated polymer membrane / about 90 wt% spherical inorganic particles, about 15 wt% fibrillated polymer membrane / about 85 wt% spherical inorganic particles, about 20 wt% fibrillated polymer membrane / about 80 wt% spherical inorganic particles, about 30 wt% fibrillated polymer membrane / about 70 wt% spherical inorganic particles, about 35 wt% fibrillated polymer membrane / about 65 wt% spherical inorganic particles, about 40 wt% fibrillated polymer membrane / about 60 wt% spherical inorganic particles, about 45 wt% fibrillated polymer membrane / about 55 wt% spherical inorganic particles, or about 50 wt% fibrillated polymer membrane / about 50 wt% spherical inorganic particles. Non-limiting examples of suitable inorganic particles include silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof. Further, the inorganic particles can be either solid or porous. Further, the affinity chromatography device described herein has a size of about 100 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ), about 150 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ), about 200 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ), about 250 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ), about 200 (×10 -12 cm 2 ) to about 450 (×10 -12 cm 2 ), about 200 (×10 -12 cm 2 ) to about 400 (×10 -12 cm 2 ), about 250 (×10 -12 cm 2 ) to about 400 (×10-12 cm 2 ) or about 300 (×10 -12 cm 2 ) to about 400 (×10 -12 cm 2 ) and has a water permeability.

[0111] In at least one embodiment, the fibrillated polymer membrane comprises a blend of spherical inorganic particles having different nominal particle sizes. For example, the fibrillated polymer membrane can comprise a 90:10 mixture of the same or different spherical inorganic particles of a first nominal particle size (e.g., 5 microns) and a second nominal particle size (e.g., 20 microns). The blend of spherical inorganic particles within the fibrillated polymer membrane can be any blend such as, for example, a 10:90 to 90:10 blend, a 30:70 to 70:30 blend, a 60:40 to 40:60 blend, a 25:75 to 75:25 blend, a 20:80 to 80:20 blend, or a 50:50 blend. In one embodiment, the inorganic particles having a spherical shape comprise 10 micron spherical particles and 20 micron spherical particles in a 10:90 to 90:10 blend. In another embodiment, the inorganic particles having a spherical shape comprise 5 micron spherical particles and 10 micron spherical particles in a 10:90 to 90:10 blend. In a further embodiment, the inorganic particles having a spherical shape comprise 5 micron spherical particles and 20 micron spherical particles in a 10:90 to 90:10 blend.

[0112] In some embodiments, the affinity ligand is covalently bound to spherical inorganic particles. In another embodiment, the affinity ligand is covalently bound to a fibrillated polymer membrane. In a further embodiment, the affinity ligand can bind to both the polymer membrane and the spherical inorganic particles. The affinity ligand can be a protein, antibody, or polysaccharide that binds reversibly to a target molecule. In one embodiment, the affinity ligand is a protein that binds reversibly, for example, to an antibody, antibody fragment, Fc fusion protein, or the Fc region of an antibody / drug conjugate. In another embodiment, the affinity ligand is an antibody, protein L, or polysaccharide that binds reversibly to a specific protein or protein fragment. Exemplary affinity ligands for use in an affinity chromatography device include, but are not limited to, protein A, protein G, protein L, human Fc receptor proteins, antibodies that specifically bind to other proteins, and heparin. The affinity ligand can be natural, recombinant, or synthetic. In yet another embodiment, the affinity ligand is a metal affinity ligand that binds reversibly to a His-tagged protein. In yet another embodiment, the affinity ligand can be an antibody or polysaccharide that binds reversibly to a specific viral vector.

[0113] In at least one embodiment, the fluoropolymer membrane is a polytetrafluoroethylene (PTFE) membrane or an expanded polytetrafluoroethylene (ePTFE) membrane. An expanded polytetrafluoroethylene (ePTFE) membrane prepared according to the method described in U.S. Patent No. 7,306,729 to Bacino et al., U.S. Patent No. 3,953,566 to Gore, U.S. Patent No. 5,476,589 to Bacino, or U.S. Patent No. 5,183,545 to Branca et al. can be used. Further, the fluoropolymer membrane can be made hydrophilic (e.g., water-wettable) using methods known in the art such as, but not limited to, the method disclosed in U.S. Patent No. 4,113,912 to Okita et al. A coating that effectively binds to a ligand as described in U.S. Patent No. 5,897,955 to Drumheller, U.S. Patent No. 5,914,182 to Drumheller, or U.S. Patent No. 8,591,932 to Drumheller can be applied to the polymer membrane.

[0114] The fluoropolymer membrane can also include a polymer material including a functional tetrafluoroethylene (TFE) copolymer membrane, where the functional TFE copolymer material includes a functional copolymer of TFE and PSVE (perfluorosulfonyl vinyl ether), or a functional copolymer of TFE and another suitable functional monomer such as, but not limited to, vinylidene fluoride (VDF), vinyl acetate, or vinyl alcohol.

[0115] Throughout this application, it is to be understood that the term "PTFE" is used herein for convenience and is intended to include not only polytetrafluoroethylene but also expanded PTFE, expanded modified PTFE, and expanded copolymers of PTFE, which are described, for example, in U.S. Patent No. 5,708,044 to Branca, U.S. Patent No. 6,541,589 to Baillie, U.S. Patent No. 7,531,611 to Sabol et al., U.S. Patent No. 8,637,144 to Ford, and U.S. Patent No. 9,139,669 to Xu et al.

[0116] Also, the fibrillated polymer membrane may be, for example, a fibrillatable polyolefin membrane (e.g., a polyethylene membrane).

[0117] The intermediate material can be a fluoropolymer film or a non-fluoropolymer film (e.g., polyethylene, expanded polyethylene, or other polyolefin films). Further, the intermediate membrane can be porous. In some embodiments, the intermediate film is a thermoplastic or thermosetting polymer film.

[0118] Advantageously, the chromatography device can be used multiple times. Further, the chromatography device can be washed with a washing solution (e.g., sodium hydroxide, phosphoric acid, citric acid, ethanol, etc.) and reused after each separation process or after multiple separation processes.

[0119] The affinity chromatography device described herein has a dynamic binding capacity (DBC) exceeding 35 mg / ml at a residence time of 20 seconds. Further, the affinity chromatography device has a cycle durability of at least 100 cycles without the operating pressure exceeding 0.3 MPa. Further, the chromatography device can be used multiple times without substantially losing its dynamic binding capacity. Specifically, the chromatography device can be washed with a washing solution (e.g., sodium hydroxide) after each separation process and reused. Embodiments of the wound membrane assembly 110 and the laminated membrane assembly 220 are described herein, but it should be understood that any number of fibrillated polymer membranes, as well as any combination of the type of fibrillated polymer membrane, the type of spherical inorganic particles, the size of the spherical inorganic particles, and the orientation of the fibrillated polymer membrane within the membrane assemblies 110, 220, are within the scope of the present disclosure. Some or all of the fibrillated polymer membranes may differ from each other in composition, thickness, permeability, etc.

[0120] The chromatographic devices and components thereof described herein can be manufactured using a variety of processes. In some embodiments, injection molding can be used to manufacture the chromatographic components provided herein. Other suitable processes include, but are not limited to, extrusion, compression molding, solvent casting, and combinations thereof. Embodiments using two fibrillated polymer membranes coextruded to produce a composite membrane assembly are also contemplated within the scope of the present disclosure. Such composite membrane assemblies can include two (or more) layers of fibrillated polymer membranes that can be coextruded or integrated together.

[0121] In some embodiments, the affinity chromatography devices described herein are utilized in a dual manifold 750 in which affinity chromatography devices 700, 701 are arranged in a parallel configuration, as generally shown in FIG. 7. It should be understood that the affinity chromatography devices shown in FIGS. 7-12 can include a laminated membrane assembly, a wound membrane assembly, or a combination thereof. As shown, an aqueous mixture flows into distribution element 740. Distribution element 740 is not particularly limited as long as it divides the flow of the aqueous mixture into at least two inlet tubes 760, 761. The divided aqueous mixture within inlet tubes 760, 761 flows into chromatography devices 700, 701 where target molecules are captured. The aqueous solution (i.e., the aqueous mixture minus the target molecules captured by the affinity ligand) flows out of chromatography devices 700, 701 through outlet tubes 780, 781, respectively. The aqueous solutions within outlet tubes 780, 781 combine within distribution element 790 and are reconstituted into a single aqueous solution. A top view of dual manifold 750 including two parallel affinity chromatography devices 700, 701 is shown in FIG. 8. In some embodiments, manifold 750 is disposed within housing 720.

[0122] Two chromatography devices 700, 701 are shown in FIGS. 7 and 8 for illustrative purposes only, and it should be understood that a plurality (i.e., three or more) of the chromatography devices described herein can be utilized in a parallel configured manifold as long as the flow and permeability distributions across the chromatography devices are similar. This similarity across the affinity chromatography devices enables scalability of device volume and performance. The affinity chromatography devices utilized in the manifold (e.g., dual, quad, etc.) may be the same as or different from each other. Further, the affinity chromatography devices can be dropped into a parallel configured system without modification or addition to the manifold 750.

[0123] An example of a manifold that includes a plurality of chromatography devices as described herein is the quad manifold shown in FIG. 9. A top view of this quad manifold 950 is shown in FIG. 10. Similar to the dual manifold shown in FIGS. 7 and 8, an aqueous mixture flows into distribution element 940. Distribution element 940 divides the flow of the aqueous mixture into inlet tubes 960, 961, 962, and 963. Note that 962 and 963 are hidden behind 961 and 962, respectively, in FIG. 9. The divided aqueous mixture within inlet tubes 960, 961, 962, and 963 (962 and 963 not shown) flows into chromatography devices 900, 901, 902, and 903 (902 and 903 not shown), respectively, where target molecules are captured. The aqueous solution flows out of each of chromatography devices 900, 901, 902, and 903 (not shown) through outlet tubes 980, 981, 982, and 983, respectively (note that 982 and 983 are hidden behind 980 and 981). The aqueous solutions within outlet tubes 980, 981, 982, and 983 (982 and 983 not shown) are combined within distribution element 990 and reconstituted into a single aqueous solution. In some embodiments, the quad manifold 950 can be disposed within housing 920.

[0124] FIG. 11 shows a device 1000 including two manifolds 1150, 1151 in a parallel configuration. A top view of the device 1000 is shown in FIG. 12. The manifolds 1150, 1151 each include four affinity chromatography devices, two of which, affinity chromatography devices 1100, 1101 and 1102, 1103 of each manifold 1150, 1151, are shown in FIG. 11. By being able to utilize at least two manifolds in parallel, the volume capacity during the use of the chromatography device described herein can be advantageously increased. In other words, the device 1000 eliminates the need to transfer to a larger volume chromatography device. Further, arranging the manifolds in parallel as shown in FIG. 11 reduces the concern of overpressurizing the device 1000.

[0125] In use, the aqueous mixture flows into distribution element 1140. Distribution element 1140 divides the flow of the aqueous mixture into two distribution tubes 1141, 1142. The divided aqueous mixture within distribution tubes 1141, 1142 is then further divided by distribution elements 1145, 1146 and introduced into the inlet tubes. In the embodiment shown in FIG. 11, inlet tubes 1160, 1161 of the four inlet tubes of manifold 1150 are shown. Similarly, inlet tubes 1162, 1163 of the four inlet tubes of manifold 1151 are shown. It should be understood that the remaining inlet tubes 1164, 1165, 1166, 1167 are hidden behind inlet tubes 1160, 1161, 1162, 1163 in FIG. 11. The aqueous mixture within the inlet tubes of the first and second manifolds 1150, 1151 flows into the affinity chromatography devices. In manifold 1150, affinity chromatography devices 1100, 1101 are shown, and in manifold 1151, affinity chromatography devices 1102, 1103 are shown. It should be understood that the remaining chromatography devices 1104, 1105, 1106, 1107 are hidden behind affinity chromatography devices 1100, 1101, 1102, 1103 in FIG. 11. The target molecules are captured by the affinity chromatography devices.

[0126] The aqueous solution flows out from each chromatography device through the outlet tubes. In FIG. 11, the outlet tubes 1180, 1181 of the manifold 1150 and the outlet tubes 1182, 1183 of the manifold 1151 are shown. It should be understood that the remaining outlet tubes are hidden behind the outlet tubes 1180, 1181, 1182, 1183 in FIG. 11. The aqueous solution in the outlet tubes is combined within the distribution elements 1170 and 1171. Next, the combined solution flows from the distribution elements 1170, 1170 into the collection tubes 1110, 1111. The aqueous solution in the collection tubes 1110, 1111 is combined at the distribution element 1190 and reconstituted into a single aqueous solution. The first manifold 1150 and the second manifold 1151 can be enclosed within the housing 1120.

[0127] In some embodiments, an affinity chromatography device within a manifold and / or the manifold itself (e.g., a dual manifold (Figure 7), a quad manifold (Figure 9)), and / or a manifold in a parallel configuration (Figure 11) can be housed within a housing. For example, there can be a first housing surrounding the chromatography device within the manifold. Thus, in a dual manifold, the first housing surrounds two affinity chromatography devices. In connection therewith, there can be a second housing surrounding the manifold (in addition to the first housing surrounding the chromatography device). In other embodiments, only the chromatography device within the manifold is surrounded by a housing. In further embodiments, the manifold and the affinity chromatography device can be housed within a single housing (e.g., the chromatography device included within the manifold is not separately housed in a housing). The material forming the housing is not particularly limited. Non-limiting examples of suitable materials include, but are not limited to, polyurethane, stainless steel, polypropylene, acrylonitrile butadiene styrene (ABS), polyethylene terephthalate (PET), polyether ether ketone (PEEK), cyclic olefin copolymer (COC), and polyethylene terephthalate glycol (PETG). Further, the shape of the housing unit is not limiting and can take any form as long as the form encapsulates the affinity chromatography device and / or the manifold.

[0128] In some embodiments, the present disclosure is directed to a diagnostic device for extracting a target substance from a biological sample. The device includes a fibrillated polymer membrane containing inorganic particles having a spherical shape and a nominal particle size of about 5 microns to about 20 microns therein. The particle size distribution has a D90 / D10 of 3 or less, and at least one of the fibrillated polymer membrane and the inorganic particles is covalently bonded to a ligand that reversibly binds to the target substance in the biological sample. The device can further include a fluid inlet and a fluid outlet fluidly connected to the fluid inlet. Further, the device can include a housing member surrounding the fluid inlet, the fluid outlet, and the fibrillated polymer membrane.

[0129] Those skilled in the art will readily understand that various aspects of the present disclosure can be implemented by any number of methods and apparatuses configured to perform the intended functions. It should also be noted that the accompanying drawings referred to herein are not necessarily drawn to scale and may be exaggerated to illustrate various aspects of the present disclosure, and in that regard, the figures should not be construed as limiting.

Examples

[0130] Test method Although specific methods and apparatuses are described below, it should be understood that other methods or apparatuses determined to be suitable by those skilled in the art can be used instead.

[0131] Method for determining the dynamic binding capacity at 10% breakthrough A chromatography device was inserted into the flow path of an AKTA™ Pure (Cytiva, Marlborough, MA) liquid chromatography system, and a single cycle consisting of the following procedure was performed. Table A shows the solutions used. Table B shows the procedure steps for determining the dynamic binding capacity at 10% breakthrough.

Table 1

Table 2

[0132] Method for determining liquid permeability The liquid permeability of the chromatography device was determined using Darcy's law. Individual devices were characterized with respect to bed cross-sectional area and bed length. Solution A was used as the liquid and characterized with respect to viscosity. The pressure drop across the column as a function of liquid flux was measured with an AKTA™ Pure liquid chromatography system.

[0133] Method for measuring particle size and particle size distribution Data on particle size and particle size distribution were provided by the manufacturer and measured using Coulter counter technology.

[0134] Purification method for CHO cell recovery The chromatography device was inserted into the flow path of an AKTA™ Pure (Cytiva, Marlborough, MA) liquid chromatography system. Table C shows the solutions used for CHO cell recovery purification. [Table 3]

[0135] Method for determining the yield of protein purification The purification yield was first determined by measuring the absorbance of the protein elution pool at a wavelength of 280 nm using a Hitachi U-2900 spectrophotometer. Next, this absorbance value was used to calculate the protein concentration of the protein elution pool using Beer's law shown below. The yield was calculated for each elution pool by dividing the total target protein mass (c) in the elution pool by the mass (c 0 ) of the loaded target protein with reference to the titration amount of either Recovery 1 or Recovery 2 from Table C using the loading volume based on Table L, M, or P. [Number]

Number

[0136] Parallel Attachment Method of Chromatography Devices All affinity chromatography devices shown in Tables K and O (described below) are attached either in a dual manifold configuration including two Y fittings and 1 / 16-inch PEEK tubing (Devices V, W, and X in Table K), or in a quad manifold configuration including six Y fittings and 1 / 16-inch Peek tubing (Device Y) or six Y fittings and 1 / 8-inch FEP tubing (Device Z), as outlined in Table D.

Table 4

[0137] Example Example 1: Laminated Membrane Reference Particles A porous polytetrafluoroethylene (ePTFE) membrane having 15 mass percent ePTFE and 85 mass percent porous silica particles with a nominal particle size of 10 microns was obtained. Further, a porous ePTFE membrane having 15 mass percent PTFE and 85 mass percent porous silica particles with a nominal particle size of 20 microns was obtained. The porous silica particles in the ePTFE membranes listed in Table E were substantially the same with respect to other chemical and physical properties such as chemical composition, particle shape, nominal particle porosity, nominal particle pore size, and nominal particle surface area. Table E shows some physical properties of the two porous ePTFE membranes.

Table 5

[0138] The porous ePTFE membranes A and B were used to manufacture an affinity chromatography device. A polypropylene flow distributor was attached to one end of a polypropylene cylinder housing. A porous polypropylene intermediate material was placed in the housing. A desired number of ePTFE membrane layers were laminated onto the polypropylene intermediate material within the housing (see Table F). A second porous polypropylene intermediate material was placed on top of the ePTFE membrane stack. A second polypropylene flow distributor was attached to the end of the cylindrical housing opposite the first polypropylene flow distributor. The chromatography device was sealed by a heating process.

[0139] The intermediate device was then treated to covalently bond Protein A to the laminated ePTFE membrane. This method is representative of methods typical to those skilled in the art and is described in more detail in U.S. Patent No. 10,525,376 to McManaway et al. and U.S. Patent No. 10,526,367 to McManaway et al.

[0140] The affinity chromatography devices manufactured as described above were tested, and their liquid permeability and dynamic binding capacity at a residence time of 20 seconds were evaluated using the procedures described in the test methods described herein. The performance of each of these affinity chromatography devices is shown in Table F.

Table 6

[0141] Example 2: Laminated Membrane - Spherical Particles with a Controlled Size Distribution Spherical particles of the size and size distribution described in Table G were obtained. The porous silica particles listed in Table G were substantially the same with respect to chemical and physical properties such as chemical composition, particle shape, nominal particle porosity, nominal particle pore size, and nominal particle surface area.

Table 7

[0142] From those in Table G, a porous polytetrafluoroethylene (ePTFE) membrane having 15 mass percent PTFE and 85 mass percent spherical porous silica particles was obtained. These ePTFE membranes had various mass mixtures of nominal particle sizes. Table H shows the mixing ratios of the respective particle sizes and the physical properties of the ePTFE membranes.

Table 8

[0143] Using the porous membranes (ePTFE) E - K, an affinity chromatography device was manufactured in the same manner as in Example 1. Next, the chromatography device was treated to covalently bond Protein A to the membrane in the same manner as in Example 1. The affinity chromatography devices were tested to evaluate their liquid permeability and dynamic binding capacity at a residence time of 20 seconds. The performance of each of these affinity chromatography devices is shown in Table I.

Table 9

[0144] The relationship between the dynamic binding capacity and liquid permeability from the devices described in Examples 1 and 2 is shown in FIG. 5 for comparison.

[0145] Example 3: Helical wound membrane Using the porous ePTFE membranes F and H described in Table H, a helical wound affinity chromatography device was constructed. Each PTFE membrane of a certain length having a porous polypropylene intermediate material at both ends was wound around a hollow polypropylene core. The length of the ePTFE membrane was sufficient to achieve the desired number of windings. The ends of the ePTFE membrane were integrated and sealed to a polypropylene distribution cap at one end and an outlet cap at the other end. The inlet cap and the outer housing were similarly sealed on the assembly such that the axially oriented channels allowed a radial flow through the ePTFE membrane layer from the outer radius to the inner radius and out through the outlet cap.

[0146] Next, in the same manner as in Example 1, each device was processed to covalently bond Protein A to the membrane. Each affinity chromatography device was tested to evaluate its liquid permeability and dynamic binding capacity at a residence time of 20 seconds. The performance of the affinity chromatography devices is shown in Table J. Figure 6 shows the relationship between devices S and T having a helical wound membrane and devices C to R having a laminated membrane.

Table 10

[0147] Example 4: Chromatography Devices V, W, and X - Dual Manifold Configuration Chromatography devices V, W, and X were constructed using the fittings and tubing outlined in Table D in a dual manifold configuration as shown in Figure 7. Five consecutive purification cycles using Recovery 1 from Table C were performed on each of chromatography devices V, W, and X. Chromatography devices V and W were carried out according to the purification conditions outlined in Table L, while chromatography device X (Cytiva, Marlborough, MA - HiScreen MabSelect SuRe LX (PN: 17547415)) was carried out according to the purification conditions outlined in Table M. Purification cycles were performed to evaluate the purification capacity and yield performance in a dual manifold configuration. Figure 13 shows a representative purification cycle for chromatography devices V and W. Figure 14 shows a representative purification cycle for chromatography device X. The yield performance of each dual manifold is shown in Table N. Chromatography devices V and W show a typical chromatogram of the purified CHO cell recovery, while chromatography device X showed evidence of early breakthrough throughout the loading step at an absorbance 500 - 600 mAU higher compared to devices V and W in Figure 13. Chromatography device X also showed evidence of a dual elution peak observable in Figure 14. This dual elution peak was not observed in devices V and W.

Table 11

Table 12

Table 13

Table 14

[0148] Example 5: Chromatography Devices Y and Z - Stacked vs. Helical Quad Manifold Configuration Chromatography devices Y and Z were configured in a quad manifold configuration as shown in FIG. 9 using the fittings and tubes outlined in Table D. A purification cycle using Recovery 2 of Table C was performed using the quad manifold. Chromatography devices Y and Z were evaluated for purification capacity and yield performance in the quad manifold configuration according to the purification conditions outlined in Table P. FIG. 15 shows representative purification cycles for both Chromatography device Y and Chromatography device Z. The yield performance of each device is shown in Table Q.

Table 15

Table 16

Table 17

[0149] The invention of this application has been described both in general and in specific embodiments. It will be apparent to those skilled in the art that various modifications and changes can be made to the embodiments without departing from the scope of the present disclosure. Therefore, the embodiments are intended to cover the modifications and variations of the present invention as long as they are within the scope of the appended claims and their equivalents. (Aspect) (Aspect 1) A fluid inlet, A fluid outlet fluidly connected to the fluid inlet, and A fibrillated polymer membrane disposed between the fluid inlet and the fluid outlet, having a spherical shape and containing inorganic particles having a nominal particle size of about 5 microns to about 20 microns therein, and A housing member surrounding the fluid inlet, the fluid outlet and the fibrillated polymer membrane, An article comprising an affinity chromatography device, The particle size distribution has a D90 / D10 of 3 or less, and at least one of the fibrillated polymer membrane and the inorganic particles is covalently bonded to an affinity ligand that reversibly binds to a target molecule. (Aspect 2) The article according to aspect 1, wherein the target molecule is a protein, an antibody, a viral vector, and combinations thereof. (Aspect 3) Having a water permeability of about 100 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ), the article according to aspect 1 or 2. (Aspect 4) The inorganic particles having a spherical shape are selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof, the article according to any one of aspects 1 to 3. (Aspect 5) The fibrillated polymer membrane comprises a stretched polytetrafluoroethylene membrane, a stretched modified polytetrafluoroethylene membrane, a stretched tetrafluoroethylene copolymer membrane, or an expanded polyethylene membrane, the article according to any one of aspects 1 to 4. (Aspect 6) The fibrillated polymer membrane is a stretched polytetrafluoroethylene membrane, the article according to any one of aspects 1 to 5. (Aspect 7) The affinity ligand is selected from protein A, protein G, protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof, the article according to any one of aspects 1 to 6. (Aspect 8) The inorganic particles include at least a first inorganic particle having a spherical shape and a first nominal particle size, and a second inorganic particle having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other, the article according to any one of aspects 1 to 7. (Aspect 9) The nominal particle size is selected from about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof, and the article according to any one of aspects 1 to 8. (Aspect 10) The inorganic particles having a spherical shape include a blend of 10-micron spherical particles and 20-micron spherical particles, and the blend is 10:90 to 90:10, and the article according to any one of aspects 1 to 9. (Aspect 11) The inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 10-micron spherical particles, and the blend is 10:90 to 90:10, and the article according to any one of aspects 1 to 9. (Aspect 12) The inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 20-micron spherical particles, and the blend is 10:90 to 90:10, and the article according to any one of aspects 1 to 9. (Aspect 13) The article according to any one of aspects 1 to 12, having a dynamic binding capacity (DBC) of at least 35 mg / ml at a residence time of 20 seconds. (Aspect 14) The article according to any one of aspects 1 to 13, having a cycle durability of at least 100 cycles at an operating pressure of less than 0.3 MPa. (Aspect 15) The fibrillated polymer film has a wound configuration, and the article according to any one of aspects 1 to 14. (Aspect 16) The fibrillated polymer film has a laminated configuration, and the article according to any one of aspects 1 to 15. (Aspect 17) The fibrillated polymer film has a wound configuration, a laminated configuration, and combinations thereof, and the article according to any one of aspects 1 to 16. (Aspect 18) The inner intermediate material is disposed around on the outer surface of the core, and the fibrillated polymer film is disposed around on the inner intermediate material, and the article according to any one of aspects 1 to 17. (Aspect 19) The outer intermediate material is disposed around on the fibrillated polymer film, and the article according to aspect 18. (Aspect 20) The inner intermediate material and the outer intermediate material are selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous nonwoven material, and a porous woven material, and the article according to aspect 18 or 19. (Aspect 21) At least one of the inner intermediate material and the outer intermediate material is a polypropylene nonwoven material, and the article according to any one of aspects 18 to 20. (Aspect 22) Use of an article according to any one of aspects 1 to 21 for separating the target molecule from a fluid stream. (Aspect 23) A manifold comprising at least two affinity chromatography devices according to any one of aspects 1 to 21 arranged in a parallel configuration. (Aspect 24) The manifold according to aspect 23, wherein the manifold is enclosed within a housing. (Aspect 25) A device comprising a first manifold and a second manifold in a parallel configuration, each of the first manifold and the second manifold comprising at least two affinity chromatography devices according to any one of aspects 1 to 21. (Aspect 26) The device according to aspect 25, wherein the first manifold and the second manifold are enclosed within a housing. (Aspect 27) A core disposed at the center, A fibrillated polymer membrane wound around the core and containing therein spherical inorganic particles having a spherical shape and a nominal particle size of from about 5 microns to about 20 microns, A housing member surrounding both the core and the fibrillated polymer membrane, A first end cap disposed at a first end of the housing member, and A second end cap disposed at a second end of the housing member, An affinity chromatography device, An article comprising, The particle size distribution has a D90 / D10 of less than 3, and at least one of the fibrillated polymer membrane and the spherical inorganic particles is covalently bonded to an affinity ligand that reversibly binds to a target molecule. (Aspect 28) The article according to aspect 27, wherein the target molecule is a protein, an antibody, a viral vector, and combinations thereof. (Aspect 29) About 100 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ) water permeability. (Aspect 30) The spherical inorganic particles having a spherical shape are selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof. (Aspect 31) The fibrillated polymer membrane comprises a stretched polytetrafluoroethylene membrane, a stretched modified polytetrafluoroethylene membrane, a stretched tetrafluoroethylene copolymer membrane, or an expanded polyethylene membrane. (Aspect 32) The fibrillated polymer film is a stretched polytetrafluoroethylene film, the article according to any one of aspects 27 to 31. (Aspect 33) The affinity ligand is selected from protein A, protein G, protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof, the article according to any one of aspects 27 to 32. (Aspect 34) The inorganic particles include at least a first inorganic particle having a spherical shape and a first nominal particle size and a second inorganic particle having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other, the article according to any one of aspects 27 to 33. (Aspect 35) The nominal particle size is selected from about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof, the article according to any one of aspects 27 to 34. (Aspect 36) The inorganic particles having a spherical shape include a blend of 10-micron spherical particles and 20-micron spherical particles, and the blend is 10:90 to 90:10, the article according to any one of aspects 27 to 35. (Aspect 37) The inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 10-micron spherical particles, and the blend is 10:90 to 90:10, the article according to any one of aspects 27 to 35. (Aspect 38) The inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 20-micron spherical particles, and the blend is 10:90 to 90:10, the article according to any one of aspects 27 to 35. (Aspect 39) Having a dynamic binding capacity (DBC) of at least 35 mg / ml at a residence time of 20 seconds, the article according to any one of aspects 27 to 38. (Aspect 40) Having a cycle durability of at least 100 cycles and an operating pressure of less than 0.3 MPa, the article according to any one of aspects 27 to 39. (Aspect 41) The inner intermediate material is disposed peripherally on the outer surface of the core, and the fibrillated polymer film is disposed peripherally on the inner intermediate material, the article according to any one of aspects 27 to 40. (Aspect 42) Including an outer intermediate material disposed peripherally on the fibrillated polymer film, the article according to any one of aspects 27 to 41. (Aspect 43) The inner intermediate material and the outer intermediate material are the article according to aspect 42, selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous nonwoven material, and a porous woven material. (Aspect 44) The article according to aspect 43, wherein at least one of the inner intermediate material and the outer intermediate material is a polypropylene nonwoven material. (Aspect 45) A manifold comprising at least two affinity chromatography devices according to any one of aspects 27 to 44 arranged in a parallel configuration. (Aspect 46) The manifold according to aspect 45, wherein the manifold is enclosed within a housing. (Aspect 47) A device comprising a first manifold and a second manifold in a parallel configuration, wherein each of the first manifold and the second manifold comprises at least two affinity chromatography devices according to any one of aspects 27 to 44. (Aspect 48) The device according to aspect 47, wherein the first manifold and the second manifold are enclosed within a housing. (Aspect 49) A housing member, An inlet allowing fluid flow into the housing member, An outlet allowing fluid flow out of the housing member, and, A laminated membrane assembly disposed within the housing member between the fluid inlet and the fluid outlet, An affinity chromatography device comprising An article comprising The laminated membrane assembly Comprises two or more fibrillated polymer membranes in a laminated configuration, each of the fibrillated polymer membranes containing therein inorganic particles having a blend of spherical and a nominal particle size of from about 5 microns to about 20 microns, The particle size distribution has a D90 / D10 of less than 3, and, The article, wherein at least one of the fibrillated polymer membrane and the spherical inorganic particles is covalently bonded to an affinity ligand that reversibly binds to a target molecule. (Aspect 50) The article according to aspect 49, comprising a first flow distributor and a second flow distributor, the first flow distributor and the second flow distributor being disposed at opposite ends of the housing member. (Aspect 51) The article according to aspect 49 or aspect 50, wherein the target molecule is a protein, an antibody, a viral vector, and combinations thereof. (Aspect 52) About 100 (×10 -12 cm 2 ) to about 500 (×10 -12 cm 2 ) of water permeability, the article according to any one of aspects 49 to 51. (Aspect 53) The inorganic particles having a spherical shape are the article according to any one of aspects 49 to 52, selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof. (Aspect 54) The fibrillated polymer film-containing article according to any one of aspects 49 to 53, wherein the fibrillated polymer film includes a stretched polytetrafluoroethylene film, a stretched modified polytetrafluoroethylene film, a stretched tetrafluoroethylene copolymer film, or an expanded polyethylene film. (Aspect 55) The fibrillated polymer film-containing article according to any one of aspects 49 to 53, wherein the fibrillated polymer film is a stretched polytetrafluoroethylene film. (Aspect 56) The article according to any one of aspects 49 to 55, wherein the affinity ligand is selected from protein A, protein G, protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof. (Aspect 57) The article according to any one of aspects 49 to 56, wherein the inorganic particles include at least first inorganic particles having a spherical shape and a first nominal particle size and second inorganic particles having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other. (Aspect 58) The article according to any one of aspects 49 to 57, wherein the nominal particle size is selected from about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof. (Aspect 59) The article according to any one of aspects 49 to 58, wherein the inorganic particles having a spherical shape include a blend of 10-micron spherical particles and 20-micron spherical particles, and the blend is from 90:10 to 10:90. (Aspect 60) The article according to any one of aspects 49 to 58, wherein the inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 10-micron spherical particles, and the blend is from 10:90 to 90:10. (Aspect 61) The article according to any one of aspects 49 to 58, wherein the inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 20-micron spherical particles, and the blend is from 10:90 to 90:10. (Aspect 62) The article according to any one of aspects 49 to 61, having a dynamic binding capacity (DBC) of at least 35 mg / ml at a residence time of 20 seconds. (Aspect 63) The article according to any one of aspects 49 to 62, having a cycle durability of at least 100 cycles and an operating pressure of less than 0.3 MPa. (Aspect 64) At least one first intermediate material is disposed on a first side of the laminate film assembly, and a second intermediate material is disposed on a second side of the laminate film assembly, the second side being opposite the first side, the article according to any one of aspects 49 to 63. (Aspect 65) The first intermediate material and the second intermediate material are selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous nonwoven material, and a porous woven material, the article according to aspect 64. (Aspect 66) At least one of the first intermediate material and the second intermediate material is a polypropylene nonwoven material, the article according to aspect 65. (Aspect 67) A manifold comprising at least two affinity chromatography devices according to any one of aspects 49 to 66 arranged in a parallel configuration. (Aspect 68) The manifold is enclosed within a housing, the manifold according to aspect 67. (Aspect 69) Comprising a first manifold and a second manifold in a parallel configuration, each of the first manifold and the second manifold comprising at least two affinity chromatography devices according to any one of aspects 49 to 66, a device. (Aspect 70) The first manifold and the second manifold are enclosed within a housing, the device according to aspect 69. (Aspect 71) Comprising a fibrillated polymer membrane containing inorganic particles having a spherical shape and a nominal particle size of from about 5 microns to about 20 microns, The particle size distribution has a D90 / D10 of 3 or less, and at least one of the fibrillated polymer membrane and the inorganic particles is covalently bonded to a ligand that reversibly binds to a target substance in a biological fluid, a diagnostic device. (Aspect 72) A fluid inlet and a fluid outlet fluidly connected to the fluid inlet, the article according to aspect 71. (Aspect 73) A housing member surrounding the fluid inlet, the fluid outlet, and the fibrillated polymer membrane, the article according to aspect 71 or 72.

Claims

1. A fluid inlet, A fluid outlet fluidly connected to the fluid inlet, and A fibrillated polymer membrane disposed between the fluid inlet and the fluid outlet and containing only inorganic particles having a spherical shape and a nominal particle size selected from about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof, and A housing member surrounding the fluid inlet, the fluid outlet, and the fibrillated polymer membrane, An article comprising an affinity chromatography device, wherein The spherical shape includes a circular or substantially circular shape in which the distance from the center of the inorganic particle to the outer edge of the particle at any point is the same distance or substantially the same distance, The particle size distribution has a D90 / D10 of 3 or less, The affinity chromatography device has a dynamic binding capacity (DBC) of at least 35 mg / ml at a residence time of 20 seconds, The affinity chromatography device has a water permeability of 100 (×10 -１２ cm ２ ) to 500 (×10 -１２ cm ２ ) and At least one of the fibrillated polymer membrane and the inorganic particles is covalently bonded to an affinity ligand that reversibly binds to a target molecule that is a protein, an antibody, a viral vector, and combinations thereof.

2. The inorganic particles having a spherical shape are selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof, according to the article of claim 1.

3. The fibrillated polymer membrane includes an expanded polytetrafluoroethylene membrane, an expanded modified polytetrafluoroethylene membrane, an expanded tetrafluoroethylene copolymer membrane, or an expanded polyethylene membrane, according to the article of any one of claims 1 to 2.

4. The fibrillated polymer membrane is an expanded polytetrafluoroethylene membrane, according to the article of any one of claims 1 to 3.

5. The affinity ligand is selected from Protein A, Protein G, Protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof, according to the article of any one of claims 1 to 4.

6. The inorganic particles include at least a first inorganic particle having a spherical shape and a first nominal particle size and a second inorganic particle having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other, according to the article of any one of claims 1 to 5.

7. The inorganic particles having a spherical shape include a blend of 10-micron spherical particles and 20-micron spherical particles, and the blend is from 10:90 to 90:10, the article according to any one of claims 1 to 6.

8. The inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 10-micron spherical particles, and the blend is from 10:90 to 90:10, the article according to any one of claims 1 to 6.

9. The inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 20-micron spherical particles, and the blend is from 10:90 to 90:10, the article according to any one of claims 1 to 6.

10. The article according to any one of claims 1 to 9, having cycle durability of at least 100 cycles at an operating pressure of less than 0.3 MPa.

11. The fibrillated polymer film has a wound configuration, the article according to any one of claims 1 to 10.

12. The fibrillated polymer film has a laminated configuration, the article according to any one of claims 1 to 11.

13. The fibrillated polymer film has a wound configuration, a laminated configuration, and combinations thereof, the article according to any one of claims 1 to 12.

14. The inner intermediate material is disposed peripherally on the outer surface of the core, and the fibrillated polymer film is disposed peripherally on the inner intermediate material, the article according to any one of claims 1 to 13.

15. The outer intermediate material is disposed peripherally on the fibrillated polymer film, the article according to claim 14.

16. The inner intermediate material and the outer intermediate material are selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous nonwoven material, and a porous woven material, the article according to claim 15.

17. At least one of the inner intermediate material and the outer intermediate material is a polypropylene nonwoven material, the article according to any one of claims 15 to 16.

18. Use of the article according to any one of claims 1 to 17 for separating the target molecule from a fluid stream.

19. A manifold including at least two affinity chromatography devices according to any one of claims 1 to 17 arranged in a parallel configuration.

20. The manifold is enclosed within a housing, the manifold according to claim 19.

21. A device comprising a first manifold and a second manifold in a parallel configuration, wherein each of the first manifold and the second manifold comprises at least two affinity chromatography devices according to any one of claims 1 to 17.

22. The device according to claim 21, wherein the first manifold and the second manifold are enclosed within a housing.

23. A core disposed at the center, A fibrillated polymer membrane wound around the core and containing only spherical inorganic particles having a nominal particle size selected from spherical and about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof, A housing member surrounding both the core and the fibrillated polymer membrane, A first end cap disposed at a first end of the housing member, and A second end cap disposed at a second end of the housing member, An affinity chromatography device comprising An article comprising The spherical shape includes a circular or substantially circular shape in which the distance from the center of the inorganic particle to the outer edge of the particle at any point is the same distance or substantially the same distance, The particle size distribution has a D90 / D10 of less than 3, The affinity chromatography device has a dynamic binding capacity (DBC) of at least 35 mg / ml at a residence time of 20 seconds, The affinity chromatography device has a water permeability of 100 (×10 -１２ cm ２ ) to 500 (×10 -１２ cm ２ ) and At least one of the fibrillated polymer membrane and the spherical inorganic particles is covalently bonded to an affinity ligand that reversibly binds to a target molecule that is a protein, antibody, virus vector, and combinations thereof.

24. The article according to claim 23, wherein the inorganic particles having a spherical shape are selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof.

25. The article according to any one of claims 23 to 24, wherein the fibrillated polymer membrane comprises a stretched polytetrafluoroethylene membrane, a stretched modified polytetrafluoroethylene membrane, a stretched tetrafluoroethylene copolymer membrane, or an expanded polyethylene membrane.

26. The article according to any one of claims 23 to 25, wherein the fibrillated polymer membrane is a stretched polytetrafluoroethylene membrane.

27. The affinity ligand is selected from protein A, protein G, protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof, and the article according to any one of claims 23 to 26.

28. The inorganic particles include at least a first inorganic particle having a spherical shape and a first nominal particle size, and a second inorganic particle having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other, and the article according to any one of claims 23 to 27.

29. The inorganic particles having a spherical shape include a blend of 10-micron spherical particles and 20-micron spherical particles, and the blend is 10:90 to 90:10, and the article according to any one of claims 23 to 28.

30. The inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 10-micron spherical particles, and the blend is 10:90 to 90:10, and the article according to any one of claims 23 to 28.

31. The inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 20-micron spherical particles, and the blend is 10:90 to 90:10, and the article according to any one of claims 23 to 28.

32. The article according to any one of claims 23 to 31, having a cycle durability of at least 100 cycles and an operating pressure of less than 0.3 MPa.

33. The inner intermediate material is disposed around on the outer surface of the core, and the fibrillated polymer film is disposed around on the inner intermediate material, and the article according to any one of claims 23 to 32.

34. The article according to claim 33, including an outer intermediate material disposed around on the fibrillated polymer film.

35. The inner intermediate material and the outer intermediate material are selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous non-woven fabric material, and a porous woven fabric material, and the article according to claim 34.

36. At least one of the inner intermediate material and the outer intermediate material is a polypropylene non-woven fabric material, and the article according to claim 35.

37. A manifold including at least two affinity chromatography devices according to any one of claims 23 to 36 arranged in a parallel configuration.

38. The manifold according to claim 37, wherein the manifold is enclosed within a housing.

39. A device comprising a first manifold and a second manifold in a parallel configuration, wherein each of the first manifold and the second manifold comprises at least two affinity chromatography devices according to any one of claims 23 to 36.

40. The device according to claim 39, wherein the first manifold and the second manifold are enclosed within a housing.

41. A housing member, An inlet that enables fluid flow to the housing member, An outlet that enables fluid flow from the housing member, and A laminated membrane assembly disposed within the housing member between the fluid inlet and the fluid outlet, An affinity chromatography device comprising An article comprising The laminated membrane assembly Comprises two or more fibrillated polymer membranes in a laminated configuration, and each of the fibrillated polymer membranes contains only inorganic particles having a nominal particle size selected from spherical and blends of about 5 microns, about 10 microns, about 15 microns, about 20 microns, and combinations thereof within, The spherical shape includes a circular or substantially circular shape in which the distance from the center of the inorganic particles to any point on the outer edge of the particles is the same distance or substantially the same distance, The particle size distribution has a D90 / D10 of less than 3, The affinity chromatography device has a dynamic binding capacity (DBC) of at least 35 mg / ml at a residence time of 20 seconds, The affinity chromatography device has a water permeability of 100 (×10 -１２ cm ２ ) to 500 (×10 -１２ cm ２ ) and, An article, wherein at least one of the fibrillated polymer membrane and the spherical inorganic particles covalently binds an affinity ligand that reversibly binds to a target molecule that is a protein, antibody, viral vector, and combinations thereof.

42. The article according to claim 41, comprising a first flow distributor and a second flow distributor, wherein the first flow distributor and the second flow distributor are disposed at both ends of the housing member.

43. The article according to any one of claims 41 to 42, wherein the inorganic particles having a spherical shape are selected from silica, zeolite, hydroxyapatite, metal oxides, and combinations thereof.

44. The fibrillated polymer film-containing article according to any one of claims 41 to 43, wherein the fibrillated polymer film comprises a stretched polytetrafluoroethylene film, a stretched modified polytetrafluoroethylene film, a stretched tetrafluoroethylene copolymer film, or an expanded polyethylene film.

45. The fibrillated polymer film-containing article according to any one of claims 41 to 43, wherein the fibrillated polymer film is a stretched polytetrafluoroethylene film.

46. The article according to any one of claims 41 to 45, wherein the affinity ligand is selected from protein A, protein G, protein L, human Fc receptor protein, antibody, polysaccharide, and combinations thereof.

47. The article according to any one of claims 41 to 46, wherein the inorganic particles at least include first inorganic particles having a spherical shape and a first nominal particle size, and second inorganic particles having a spherical shape and a second nominal particle size, and the first nominal particle size and the second nominal particle size are different from each other.

48. The article according to any one of claims 41 to 47, wherein the inorganic particles having a spherical shape include a blend of 10-micron spherical particles and 20-micron spherical particles, and the blend is from 90:10 to 10:

90.

49. The article according to any one of claims 41 to 47, wherein the inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 10-micron spherical particles, and the blend is from 10:90 to 90:

10.

50. The article according to any one of claims 41 to 47, wherein the inorganic particles having a spherical shape include a blend of 5-micron spherical particles and 20-micron spherical particles, and the blend is from 10:90 to 90:

10.

51. The article according to any one of claims 41 to 50, having a cycle durability of at least 100 cycles and an operating pressure of less than 0.3 MPa.

52. The article according to any one of claims 41 to 51, wherein at least one first intermediate material is disposed on a first side of the laminated film assembly, and a second intermediate material is disposed on a second side of the laminated film assembly, and the second side is opposite to the first side.

53. The article according to claim 52, wherein the first intermediate material and the second intermediate material are selected from a porous fluoropolymer film, a porous non-fluoropolymer film, a porous nonwoven material, and a porous woven material.

54. The article according to claim 53, wherein at least one of the first intermediate material and the second intermediate material is a polypropylene nonwoven fabric material.

55. A manifold comprising at least two affinity chromatography devices according to any one of claims 41 to 54 arranged in a parallel configuration.

56. The manifold according to claim 55, wherein the manifold is enclosed within a housing.

57. A device comprising a first manifold and a second manifold in a parallel configuration, each of the first manifold and the second manifold comprising at least two affinity chromatography devices according to any one of claims 41 to 54.

58. The device according to claim 57, wherein the first manifold and the second manifold are enclosed within a housing.

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